CN110452857A - One plant of lactobacillus plantarum for producing non-protein micromolecular antibacterial metabolin and its application - Google Patents
One plant of lactobacillus plantarum for producing non-protein micromolecular antibacterial metabolin and its application Download PDFInfo
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- CN110452857A CN110452857A CN201910843322.5A CN201910843322A CN110452857A CN 110452857 A CN110452857 A CN 110452857A CN 201910843322 A CN201910843322 A CN 201910843322A CN 110452857 A CN110452857 A CN 110452857A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12R2001/00—Microorganisms ; Processes using microorganisms
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- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
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Abstract
The invention discloses the lactobacillus plantarum KZ0310 that deposit number is CGMCC No.17712, thallus and its metabolin have broad spectrum antibiotic activity, strong bacteriostasis is shown to a variety of gram-positive bacterias and Gram-negative bacteria (including pathogenic bacteria and spoilage organisms), it can be used for preparing antibacterial or/and aseptic production, applied to fields such as medical treatment, food to control bacterial pathogen or spoilage organisms.Antibacterial metabolin of the invention is different from bacteriocin lab or antibacterial peptide, for non-protein micromolecular substance, it can inactivate, be played to be conducive to stablizing for antibacterial activity, with good development and application prospects to avoid by people, the intracorporal proteasome degradation of animals and plants when in use.
Description
Technical field
The invention belongs to microorganisms technical fields, are related to the plant cream bar of one plant of production non-protein micromolecular antibacterial metabolin
Bacterium and its application.
Background technique
Antibacterial agent including antibiotic has not only saved the life of countless people, and help people extend the service life and
Improve quality of life.Unfortunately, be excessively used and misapply antibacterial agent (including non-treatment purpose animal, food,
Improper use in agricultural and aquatic products industry) it results in the appearance of resistant microorganism and largely propagates.Antibacterial agent resistance is all micro-
It is found in biology, either bacterium, fungi, virus or helminth.With the fast development of global trade and tourism, resistance
Microorganism can travel to any one global corner.However, current antibacterials status is: on the one hand, existing
Drug is not able to satisfy the needs for the treatment of；On the other hand, without antibacterials new enough be developed replace it is old or by
The drug gradually to fail.As a result, many common infection gradually become dangerous or can not treat, so as to cause longer treatment cycle
With the higher death rate, epoch as returned to not antibiotic.Antibacterial agent resistance has become the significant threat of human health
One of, therefore, it is badly in need of developing new antibacterials or antibacterial means effectively to kill pathogenic bacteria.
Lactic acid bacteria is a kind of anaerobism or amphimicrobian Gram-positive bacillus or coccus, does not form gemma, do not move, nothing
Cytochromes and catalase activity, glucose fermentation can be generated 50% or more lactic acid bacterium general name.Lactic acid bacteria quilt
FDA is considered the microorganism with GRAS security level, distributes widely in nature, including animals and plants body surface, dairy products, hair
In ferment food etc..Lactic acid bacteria can generate the metabolin for being conducive to food preservation, such as CO2, biacetyl, hydrogen peroxide, fatty acid, cream
Acid, benzoic acid, bacteriocin, Cyclic dipeptides etc., people often process and handle certain food using the characteristic of lactic acid bacteria metabolite,
It improves the palatability of food or extends storage phase.
Summary of the invention
One of the objects of the present invention is to provide a strains of lactic acid bacteria, thallus and its metabolin have broad spectrum antibiotic activity；Mesh
Two be to provide the preparation method of the antibacterial metabolin of the lactic acid bacteria；The third purpose is to provide the lactic acid bacteria and its antibacterial
The purposes of metabolin.
Through studying, the invention provides the following technical scheme:
Lactobacillus plantarum 1. (Lactobacillus plantarum) KZ0310, deposit number is CGMCC No.17712.
Lactobacillus plantarum KZ0310 of the invention is to separate to obtain from the pickles water of Homemade traditional zymotic, in 2019
On May 6, in is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC, address: Beijing
The institute 3 of city, North Star West Road, Chaoyang District 1), deposit number is CGMCC No.17712, and classification naming is lactobacillus plantarum
(Lactobacillus plantarum)。
2. lactobacillus plantarum KZ0310 or its bacteria suspension or its fermentation liquid or its metabolin are produced in preparation antibacterial or/and anti-corrosion
Application in product.
Further, the antibacterial or/and aseptic production inhibit gram-positive bacteria and Gram-negative bacteria simultaneously.
Further, the antibacterial or/and aseptic production inhibit escherichia coli (Escherichia coli), verdigris false
Monad (Pseudomonas aeruginosa), salmonella (Salmonella), the rugged Cronobacter enterobacteria of slope
(Enterobacter sakazakii), Listeria monocytogenes (Listeria monocytogenes), golden yellow
Any one of staphylococcus (Staphylococcus aureus) and enterococcus faecalis (Enterococcus faecalis) or
It is a variety of.
Wherein, the rugged Cronobacter enterobacteria of escherichia coli, pseudomonas aeruginosa, salmonella, slope is Gram-negative
Bacterium, Listeria monocytogenes, staphylococcus aureus and enterococcus faecalis are gram-positive bacteria.
3. the antibacterial metabolin generated by lactobacillus plantarum KZ0310.
Further, the antibacterial metabolin is non-protein micromolecular substance, and molecular weight is lower than 500Da.
4. the preparation method of the antibacterial metabolin, comprising the following steps:
(1) lactobacillus plantarum KZ0310 is inoculated in stationary culture in MRS fluid nutrient medium, gained fermentation liquid acetic acid second
Ester extraction, collects middle layer, and rotary evaporation removes ethyl acetate, obtains metabolin study；
(2) the metabolin study that step (1) obtains first is subjected to decoloration removal of impurities with nonpolar macroporous adsorption resin, then carried out
Chromatographic separation and purification carries out Determination of Antibacterial Activity finally to get antibacterial metabolin is arrived.
Further, the step (1) is that lactobacillus plantarum KZ0310 is inoculated in MRS liquid with the inoculum concentration of 0.1wt%
In culture medium, 30 DEG C of stationary culture 72h, gained fermentation liquid is extracted with ethyl acetate, and collects middle layer, and rotary evaporation removes acetic acid
Ethyl ester obtains metabolin study.
Further, the step (2) is that nonpolar macroporous absorption tree is added in the metabolin study that step (1) obtains
Rouge, 4-16 DEG C, 200rpm shake cleaned with decolourizing overnight, then successively use gel filtration chromatography, reverse-phase chromatography and high-efficient liquid phase color
Spectrum is isolated and purified, and is collected separating liquid and is carried out Determination of Antibacterial Activity to get antibacterial metabolin is arrived.
5. application of the antibacterial metabolin in preparation antibacterial or/and aseptic production.
Further, the antibacterial or/and aseptic production inhibit gram-positive bacteria and Gram-negative bacteria simultaneously.
Further, the antibacterial or/and aseptic production inhibit escherichia coli (Escherichia coli), verdigris false
Monad (Pseudomonas aeruginosa), salmonella (Salmonella), the rugged Cronobacter enterobacteria of slope
(Enterobacter sakazakii), Listeria monocytogenes (Listeria monocytogenes), golden yellow
Any one of staphylococcus (Staphylococcus aureus) and enterococcus faecalis (Enterococcus faecalis) or
It is a variety of.
The beneficial effects of the present invention are: the present invention provides a kind of lactobacillus plantarum KZ0310, thallus and its metabolin
With broad spectrum antibiotic activity, a variety of gram-positive bacterias and Gram-negative bacteria (including pathogenic bacteria and spoilage organisms) are shown
Strong bacteriostasis can be used for preparing antibacterial or/and aseptic production, applied to fields such as medical treatment, food to control bacterial pathogen
Bacterium or spoilage organisms.Antibacterial metabolin of the invention is different from bacteriocin lab or antibacterial peptide, is non-protein micromolecular substance,
It can inactivate when in use to avoid by people, the intracorporal proteasome degradation of animals and plants, be played to be conducive to stablizing for antibacterial activity,
With good development and application prospects.
Detailed description of the invention
Fig. 1 is lactobacillus plantarum KZ0310 thallus to the bacteriostatic activity of pathogenic bacteria, and wherein S is staphylococcus aureus, and E is
Escherichia coli.
Fig. 2 is bacteriostatic activity of the lactobacillus plantarum KZ0310 thallus to 7 kinds of pathogenic bacteria, wherein marking different English alphabets
There are significant difference (P < 0.05) between the data of (a, b, c).
Fig. 3 is that the resin decolorization removal of impurities of metabolin study is handled, and wherein A is the color of decoloration removal of impurities front and back sample, and S is de-
For color removal of impurities front and back sample to the bacteriostatic activity of staphylococcus aureus, E is decoloration removal of impurities front and back sample to escherichia coli
Bacteriostatic activity.
Fig. 4 is gel filtration chromatography separation, and wherein A is Superdex 30increase 10/30GL solvent resistant column point
From curve, curve 1 is UV absorption curve, and curve 2 is conductance cond curve；B is that 6 component of pipe 5 and pipe (peak shown in arrow in A) is right
The bacteriostatic activity of staphylococcus aureus.
Fig. 5 is reverse-phase chromatography (RPC) separation, and wherein A is 3mL reverse phase post separation curve, and curve 1 is UV absorption curve, bent
Line 2 is gradient elution curve；B is pipe 4,7 component of pipe 5, pipe 6 and pipe (part shown in arrow in A) to staphylococcus aureus
Bacteriostatic activity.
Fig. 6 is high performance liquid chromatography (HPLC) separation, and wherein A is that HPLC separates curve, and B is at retention time 3.143min
Peak component (peak shown in arrow in A) to the bacteriostatic activity of staphylococcus aureus.
Fig. 7 is the protease stability of antibacterial metabolin.
Specific embodiment
To make the objectives, technical solutions, and advantages of the present invention clearer, below in conjunction with attached drawing to of the invention excellent
Embodiment is selected to be described in detail.
Embodiment 1: the screening and identification of antibacterial lactic acid bacteria
(1) separation of microbial strains
Pickles water sample is diluted with 10 times of dilution gradient with sterile saline.100 μ L 10 are taken respectively-2、
10-3、10-4、10-5The pickles water diluent of dilution is applied on MRS solid medium, 37 DEG C of constant temperature incubation 48h.Picking cream
Sour bacterium single colonie further repeatedly isolates and purifies feature bacterium colony using scribing line partition method, records the form of each bacterium colony
Feature simultaneously uses gram stain microscopy, observation.
The MRS solid medium consists of the following compositions based on mass percentage: glucose 2%, peptone 1%,
Beef extract 1%, yeast extract 0.5%, sodium acetate 0.5%, Tween 80 0.1%, citric acid hydrogen diamine 0.1%, magnesium sulfate
0.02%, manganese sulfate 0.005%, agar powder 1.5%, surplus are distilled water.
KZ0310 bacterial strain colony morphology characteristic: colony shape is round, surface is smooth and slightly convex on MRS agar plate
It rises, colony colour is light milky white.
KZ0310 bacterial strain individual morphology feature: Gram-positive (G+), rod-short is single or pairs of, no gemma, no whip
Hair.
(2) screening of antibacterial lactic acid bacteria
The screening of antibacterial lactic acid bacteria is carried out using the double-deck method of scoring.Lactobacillus inoculum is cultivated into MRS fluid nutrient medium
To OD600nm=0.8, bacteria suspension is crossed on MRS agar plate (2 parallel lines, length about 2cm), 37 DEG C of perseverances with oese
Temperature culture 12h.10mL is contained 107CFU/mL indicator bacteria (staphylococcus aureus ATCC25923, escherichia coli
ATCC25922 LB agar medium (0.7wt% agar)) is poured into above-mentioned MRS media surface, 37 DEG C of constant temperature incubations for 24 hours,
Observe inhibition zone size.
As a result as shown in Figure 1, KZ0310 bacterial strain staphylococcus aureus and escherichia coli are shown it is strong antibacterial
Circle.
(3) identification of microbial strains:
By KZ0310 strain inoculated in MRS fluid nutrient medium, 37 DEG C of culture 12h.It pressesGenomic DNA
Mini Kit (Invitrogen, USA) kit specification method extracts lactic acid bacteria genomic DNA.With lactic acid bacteria genomic DNA
For template, 27F (SEQ ID No.1) and 1495R (SEQ ID No.2) are primer progress 16s rDNA amplification, and to PCR amplification
Product carries out sequencing analysis.
The 16s rDNA sequence of the KZ0310 bacterial strain of acquisition carries out in ncbi database as shown in SEQ ID No.3
BLAST compares analysis, the results show that KZ0310 bacterial strain and lactobacillus plantarum (Lactobacillus plantarum)
Identity=99% (> 97%).Therefore, KZ0310 bacterial strain is lactobacillus plantarum (Lactobacillus plantarum), will
It is named as KZ0310, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on May 6th, 2019
(abbreviation CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3), deposit number are CGMCC No.17712.
Embodiment 2: the antibacterial activity of lactobacillus plantarum KZ0310
By enterococcus faecalis, escherichia coli, staphylococcus aureus, Listeria monocytogenes, salmonella,
7 kinds of pathogenic bacteria of the rugged Cronobacter enterobacteria of slope and pseudomonas aeruginosa are activated.Each pathogenic bacteria inoculation that 0.1mL is incubated overnight
Into 100mL fresh LB agar medium (0.7wt% agar), mixes, use bilayer method of scoring described in embodiment 1 (2)
Lactobacillus plantarum KZ0310 is measured to the antibacterial circle diameter of this 7 kinds of pathogenic bacteria (perpendicular to bacterium line direction calculating).
As a result as shown in Fig. 2, lactobacillus plantarum KZ0310 shows good bacteriostatic activity to 7 kinds of pathogenic bacteria, suppression
The strong and weak sequence of bacterium ability is successively are as follows: and Listeria monocytogenes > escherichia coli > pseudomonas aeruginosa > salmonella >
The rugged Cronobacter enterobacteria > staphylococcus aureus > enterococcus faecalis of slope.It can be seen that lactobacillus plantarum KZ0310 is anti-with wide spectrum
Bacterium activity, is able to suppress various pathogens, can be used for preparing antibacterials, is drawn for treating by following any one or more bacteriums
The infectious diseases risen: Listeria Monocytogenes, escherichia coli, pseudomonas aeruginosa, salmonella, slope are rugged
Cronobacter enterobacteria, staphylococcus aureus and enterococcus faecalis；It can also be by lactobacillus plantarum KZ0310 activity thallus or its bacterium
Suspension is used to prepare preservative, is added in food, drug etc., inhibits the growth and breeding of microorganism, prevents protected object corrupt
It is rotten.
Embodiment 3: extraction, partial purification and the antibacterial activity of antibacterial metabolin
(1) extraction of antibacterial metabolin
Picking lactobacillus plantarum KZ0310 single colonie is inoculated in 10mL MRS fluid nutrient medium, 37 DEG C, 150rpm it is overnight
Culture, as kind of a daughter bacteria.Kind of a daughter bacteria is inoculated into 1L MRS fluid nutrient medium with the inoculum concentration of 0.1wt%, 30 DEG C of standing trainings
72h is supported, is centrifuged (8000rpm, 15min, 4 DEG C), supernatant is extracted with ethyl acetate, and collects the milky white liquid of middle layer, rotary evaporation
Ethyl acetate is removed, lactobacillus plantarum KZ0310 metabolin study is obtained.
Enterococcus faecalis, escherichia coli, staphylococcus aureus, monocyte are increased using Odontothrips loti measurement study
Give birth to the bacteriostatic activity of Listeria, salmonella, slope rugged Cronobacter enterobacteria and 7 kinds of pathogenic bacteria of pseudomonas aeruginosa.As a result it shows
Show, the metabolin study of lactobacillus plantarum KZ0310 all has bacteriostatic activity to 7 kinds of pathogenic bacteria, and antibacterial circle diameter size exists
Between 17.7-25.1mm.
(2) partial purification of antibacterial metabolin
20g nonpolar macroporous adsorption resin, 16 DEG C, 200rpm mistake are added in lactobacillus plantarum KZ0310 metabolin study
Night concussion, with adsorpting pigment and part protein matter.The color and sample of decoloration removal of impurities front and back sample are (golden yellow to indicator bacteria
Staphylococcus A TCC25923, escherichia coli ATCC25922) bacteriostatic activity it is as shown in Figure 3.After resin treatment, sample
Color become light yellow from dark brown black.
Sample after decoloration removal of impurities is separated using Superdex 30increase 10/30GL solvent resistant column, is pressed
1mL/ pipe collects separating liquid, and carrying out Determination of Antibacterial Activity to each pipe collection liquid (is instruction with staphylococcus aureus ATCC25923
Bacterium).As shown in figure 4, having obtained 7 through Superdex 30increase 10/30GL gel filtration post separation and significantly having divided
From peak, wherein peak (in figure arrow shown in peak) component of the retention volume in the section 23-25mL has bacteriostatic activity.The splitter
Column volume is 24mL, and the time of antibacterial activity peak appearance occurs after the main peak of Yan Feng (according to cond value), it is possible thereby to judge,
The group at the antibacterial activity peak is divided into small-molecule substance, and molecular weight is lower than 500Da.
The isolated antibacterial activity peak component of Superdex 30increase 10/30GL solvent resistant column is closed
And after vacuum refrigeration concentration, then separated using RPC 3mL reversed-phase column, collects separating liquid by 0.5mL/ pipe, each pipe is received
Liquid collecting carries out Determination of Antibacterial Activity (using staphylococcus aureus ATCC25923 as indicator bacteria).As shown in figure 5, anti-through RPC 3mL
Phase post separation has obtained 8 obvious detached peaks, wherein peak (in figure arrow shown in peak) of the retention volume in the section 4-5mL
Component has bacteriostatic activity.
Finally, the isolated antibacterial activity peak component of RPC 3mL reversed-phase column is merged, after vacuum refrigeration concentration,
It is separated using HPLC ZORBAX 300Extend-C18 column, is collected by appearance, it is living to carry out antibacterial to each peak collection liquid
Property measurement (using staphylococcus aureus ATCC25923 as indicator bacteria).As shown in fig. 6, separated through HPLC, obtained 5 it is brighter
Aobvious detached peaks, wherein peak (in figure arrow shown in peak) component of the retention time at 3.143min has bacteriostatic activity.
(3) Protease Treatment
By the antibacterial activity sample of HPLC purified pool, 4h, 100 DEG C of water are handled using Proteinase K (proteinase K)
After bath heating enzyme inactivation processing, the remaining antibacterial activity that Odontothrips loti measures sample (is to refer to escherichia coli ATCC25922
Show bacterium), replace Proteinase K to carry out same treatment as control (control) using isometric buffer.As a result as shown in fig. 7, through
Sample after Protease Treatment, antibacterial activity do not have any loss, illustrate that the small-molecule substance with antibacterial activity is non-protein
Substance.
Finally, it is stated that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although passing through ginseng
According to the preferred embodiment of the present invention, invention has been described, it should be appreciated by those of ordinary skill in the art that can
To make various changes to it in the form and details, without departing from the present invention defined by the appended claims
Spirit and scope.
Sequence table
<110>Southwest University
<120>one plants of lactobacillus plantarums for producing non-protein micromolecular antibacterial metabolin and its application
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 1
agagtttgat cctggctcag 20
<210> 2
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
ctacggctac cttgttacga 20
<210> 3
<211> 1048
<212> DNA
<213>lactobacillus plantarum KZ0310 (Lactobacillus plantarum KZ0310)
<400> 3
gtgcaggcgg cgtgctatac atgcaagtcg aacgaactct ggtattgatt ggtgcttgca 60
tcatgattta catttgagtg agtggcgaac tggtgagtaa cacgtgggaa acctgcccag 120
aagcggggga taacacctgg aaacagatgc taataccgca taacaacttg gaccgcatgg 180
tccgagtttg aaagatggct tcggctatca cttttggatg gtcccgcggc gtattagcta 240
gatggtgggg taacggctca ccatggcaat gatacgtagc cgacctgaga gggtaatcgg 300
ccacattggg actgagacac ggcccaaact cctacgggag gcagcagtag ggaatcttcc 360
acaatggacg aaagtctgat ggagcaacgc cgcgtgagtg aagaagggtt tcggctcgta 420
aaactctgtt gttaaagaag aacatatctg agagtaactg ttcaggtatt gacggtattt 480
aaccagaaag ccacggctaa ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg 540
ttgtccggat ttattgggcg taaagcgagc gcaggcggtt ttttaagtct gatgtgaaag 600
ccttcggctc aaccgaagaa gtgcatcgga aactgggaaa cttgagtgca gaagaggaca 660
gtggaactcc atgtgtagcg gtgaaatgcg tagatatatg gaagaacacc agtggcgaag 720
gcggctgtct ggtctgtaac tgacgctgag gctcgaaagt atgggtagca aacaggatta 780
gataccctgg tagtccatac cgtaaacgat gaatgctaag tgttggaggg tttccgccct 840
tcagtgctgc agctaacgca ttaagcattc cgcctgggga gtacggccgc aaggctgaaa 900
ctcaaaggaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgaagcta 960
cgcgaagaac cttaccaggt cttgacatac tatgcaaatc taagagatta gacgtttcct 1020
tcggggacat ggatacaggt ggtgcatg 1048
Claims (10)
- Lactobacillus plantarum 1. (Lactobacillus plantarum) KZ0310, deposit number is CGMCC No.17712.
- 2. lactobacillus plantarum KZ0310 described in claim 1 or its bacteria suspension or its fermentation liquid or its metabolin are in preparation antibacterial Or/and the application in aseptic production.
- 3. application as claimed in claim 2, which is characterized in that the antibacterial or/and aseptic production inhibit gram positive simultaneously Property bacterium and Gram-negative bacteria.
- 4. the antibacterial metabolin that the lactobacillus plantarum KZ0310 as described in claim 1 is generated.
- 5. antibacterial metabolin as claimed in claim 4, which is characterized in that be non-protein micromolecular substance, molecular weight is lower than 500Da。
- 6. the preparation method of antibacterial metabolin described in claim 4, which comprises the following steps:(1) lactobacillus plantarum KZ0310 is inoculated in stationary culture in MRS fluid nutrient medium, gained fermentation liquid is extracted with ethyl acetate It takes, collects middle layer, remove ethyl acetate, obtain metabolin study；(2) the metabolin study that step (1) obtains first is subjected to decoloration removal of impurities with nonpolar macroporous adsorption resin, then carries out chromatography It isolates and purifies, and carries out Determination of Antibacterial Activity to get antibacterial metabolin is arrived.
- 7. the preparation method of antibacterial metabolin as claimed in claim 6, which is characterized in that the step (1) is with 0.1wt% Lactobacillus plantarum KZ0310 is inoculated in MRS fluid nutrient medium by inoculum concentration, 30 DEG C of stationary culture 72h, gained fermentation liquid second Acetoacetic ester extraction, collects middle layer, and rotary evaporation removes ethyl acetate, obtains metabolin study.
- 8. the preparation method of antibacterial metabolin as claimed in claim 6, which is characterized in that the step (2) is obtained in step (1) To metabolin study in be added nonpolar macroporous adsorption resin, 4-16 DEG C, 200rpm shake cleaned with decolourizing overnight, then successively It is isolated and purified with gel filtration chromatography, reverse-phase chromatography and high performance liquid chromatography, collect separating liquid and carries out antibacterial activity survey Determine to get antibacterial metabolin is arrived.
- 9. application of the antibacterial metabolin described in claim 4 in preparation antibacterial or/and aseptic production.
- 10. application as claimed in claim 9, which is characterized in that the antibacterial or/and aseptic production inhibit gram positive simultaneously Property bacterium and Gram-negative bacteria.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110408577A (en) * | 2019-09-06 | 2019-11-05 | 西南大学 | One plant of Lactobacillus casei for controlling vegetables bacterial soft rot and its application |
| CN115851491A (en) * | 2022-08-31 | 2023-03-28 | 西北农林科技大学 | Lactobacillus plantarum and application thereof |
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| CN115851491A (en) * | 2022-08-31 | 2023-03-28 | 西北农林科技大学 | Lactobacillus plantarum and application thereof |
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